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1 INTRODUCTION TO DOCTORAL THESIS Necessity of the thesis Tuberculosis (TB) is a chronic infection caused by Mycobacterium tuberculosis TB bacterium was discovered by Robert Koch in 1882 and TB is the leading cause of death in infectious diseases Mycobacterium tuberculosis can be found in all parts of the body, and pulmonary tuberculosis is the most common form of TB (accounting for 80-85%) and is the main source of infection around the world Vietnam is still a country with a relatively high burden of tuberculosis, ranking 16th among 30 countries with the highest number of tuberculosis patients in the world and ranking 15th out of 30 countries with the highest burden of multi-drug resistant TB gender The total number of newly diagnosed TB cases (including HIV+) in 2017 is 124.000 people; Percentage of detection of tuberculosis is 129 per 100.000 people, in which the rate of multi-drug resistance in new patients (%) is 4.1/100.000 people; Annual TB death (excluding HIV) is 12.000 people Objectives 1) To study laboratory characteristics of peripheral blood and bone marrow in patients with non-drug-resistant pulmonary tuberculosis treated in the National Lung Hospital from 2015 to 2017 2) To evaluate changes in hematological characteristics in patients before and after treatment with anti-tuberculosis drugs Practical impact and scientific contribution of the thesis 3.1 New scientific contributions: - The research results showed the proportion of people with pulmonary tuberculosis having hematological disorders; the results also show some changes related to disease stage: treatment and untreated; and changes after one month of treatment - The research results showed a high rate of anemia in patients with pulmonary tuberculosis (over 70%), changes in white blood cells and platelets counts and morphological characteristics Regarding hematopoietic bone marrow the research results showed the status of changes in bone marrow cells as well as hematopoietic features and common secondary hematopoietic disorders - The research results of the dissertation showed a picture of blood and bone marrow features in patients with pulmonary tuberculosis, such as a direct bone marrow abnormalities due to bacterial infection, and the ability of recovery of bone marrow after anti-tuberculosis treatment 3.2 Practical impact of the thesis: - The results of the thesis are highly applicable, contributing to predicting the causes of disorders in order to help clinicians with options of diagnosis, prognosis and treatment of pulmonary tuberculosis - The results of the thesis showed a significant association between hematological indicators in certain types of pulmonary tuberculosis - The research results have shown a clear effect on positive changes in posttreatment indicators such as improvement of anemia, recovery of secondary bone marrow diseases in patients with pulmonary tuberculosis Structure of the thesis - The thesis consists of 123 pages, including: introduction - pages, literature review - 29 pages, subjects and research methodology - 11 pages, results - 40 pages, discussion - 38 pages, conclusion - pages and recommendations - page - The thesis consists of 35 tables, 29 charts, diagram and 54 images There were 146 references, from which 125 in English, 21 in Vietnamese Majority of references was published within recent 10 years The appendix includes references, lists of patients with non-drug-resistant pulmonary tuberculosis, images illustrating blood tests features, histograms, images of test results of cytology, coagulation, biochemistry and immunology Chapter 1: LITERATURE OVERVIEW 1.1 PULMONARY TUBERCULOSIS (PTB) 1.1.1 Pathogen Tuberculosis caused by Mycobacterium tuberculosis, discovered by Robert Koch in 1882, is also called Bacilie de Koch (abbreviated as BK) TB bacterium belongs to the Mycobacterium family Mycobacteria are aerobic bacterium, often with a slightly curved, non-mobile bacilli, size 0,2µm-0,6µm x 1,0µm-10µm Ziehl-Neelsen dyes catches red on a green background, free from alcohol and acids, which discolors fucsin, hence they also called acid fast bacilli (AFB) Based on this feature, it is possible to detect TB bacteria in specimens by screening AFB 1.2.2 Location of infection TB bacteria can enter the body through many entry sites, mainly through the respiratory tract, but also can be through the gastrointestinal tract, skin, conjunctiva After causing primary damage, TB bacteria can spread through lymphatic or blood stream to other organs causing secondary damage Many organs such as lungs, kidney, meninges, bones, skin, lymph nodes can be infected with tuberculosis, but most often are the lungs (80-85%), the common infective location in the lungs is the apical lobe, because of slow blood flow and bacteria can proliferate to cause an active disease 1.2.3 Diagnostic criteria Diagnostic criteria are based on the result of sputum smear, which directly showed AFB: a AFB (+) pulmonary tuberculosis: has at least sputum sample or bronchial fluid, gastric fluid with direct AFB(+) results at laboratories accredited by the National TB Program b AFB (-) pulmonary tuberculosis: when there are at least samples of AFB sputum (-),the procedure in patients is needed to diagnose AFB pulmonary TB (-) Patients who are diagnosed with AFB pulmonary tuberculosis (-) should satisfy of the following conditions: - There is evidence of tuberculosis in sputum, bronchial fluid, and gastric juice by culture method or new techniques such as Xpert MTB / RIF - Final diagnosis is made by a physician and a complete TB treatment regimen is based on: (1) clinical symptoms, (2) abnormal tuberculosis suspected loci on chest radiographs and (3) add of the following criteria: HIV (+) or not responding to broad spectrum antibiotic treatment 1.2.4 Classification of tuberculosis 1.2.4.1 Newly diagnosed TB 1.2.4.2 Relapsed TB 1.2.4.3 Treatment failure TB 1.2.4.4 Re-treatment after incompliance 1.2.4.5 Other 1.2.4.6 Transferred patients 1.2.5 Indications and treatment regimens The National Tuberculosis Program provides five of essential anti-tuberculosis drugs line 1: isoniazide (INH), rifampicin (RMP), pyrazinamide (PZA), ethambutol (E) and streptomycin (S) 1.2.5.1 Regimen I: 2RHZE/4RHE or 2RHZS/4RHE Indication: For new cases of TB (no treatment for TB ever or for TB treatment for less than one month) 4 1.2.5.2 Regimen II: 2SRHZE/1RHZE/5RHE or 2SRHZE/1RHZE/5R3H3E3 Indications: For cases of recurrent TB, failure, re-treatment after giving up and tuberculosis schools are classified as "different" 1.2 CHARACTERISTICS SOME HEMATOLOGICAL INDICES IN PTB 1.2.1 Red blood cells: Pulmonary tuberculosis reduces the number of erythrocytes, decreases the hemoglobin concentration due to tuberculosis infection, and changes the chemistry and the characteristics of erythrocyte membranes, leading to a decrease in the elasticity of erythrocytes, increased erythrocyte agglutination, and reduced ability move of red blood cells in circulation, especially in microchip, thereby reducing the ability to transport oxygen to organs 1.2.2 Leukocytes: Pulmonary TB can cause an increase in the number of leukocytes, neutrophilia, monocytosis, acidophilia and sometimes cause lymphopenia Changes in the number and function of white blood cells are confirmed in all cases of infection, especially in patients with latent TB or active pulmonary tuberculosis 1.2.3 Platelet: Platelet count usually increases in about 52% in people with tuberculosis Normal platelet counts are also one of the signs that treatment has been successful In people with pulmonary tuberculosis accompanied by reduced birth marrow or phagocytic syndrome, platelet counts are often severely reduced; sometimes very heavy 1.2.4 Bone marrow characteristics in pulmonary tuberculosis: Secondary bone marrow diseases are common in combination disease of pulmonary tuberculosis, sometimes only red blood cells or platelets, sometimes disturbing both red blood cells and platelets Chapter 2: STUDY SUBJECTS AND METHODS 2.1 STUDY SUBJECTS: Including 158 patients who agreed to participate in the study and were diagnosed as non-drug-resistant pulmonary tuberculosis according to the standards of the National TB Program Grouping patients in research subjects: - Patients with pulmonary tuberculosis non-drug-resistant to study the characteristics of peripheral blood and bone marrow testing of 158 patients Include: + The group of patients with new pulmonary tuberculosis non-drug-resistant (formula I) is 111 patients + The group of patients with pulmonary tuberculosis treated non-drug-resistant (formula II) was 47 patients - The group of patients with non-drug-resistant pulmonary tuberculosis tested for the second time after month of anti-tuberculosis treatment was 33 patients 2.2 STUDY TIME AND LOCATION 158 patients with pulmonary 2.2.1 Time: From March 2015 to July 2017 tuberculosis non-drug-resistant are 2.2.2 Location: National Lung Hospital prescribed anti-TB drugs 2.2.3 Patient selection criteria: (111 patients with new PTB and 47 - Inpatient over 16 years old patients PTB treated) - Patients are diagnosed with TB non-drug-resistant and are treated with antituberculosis drugs line (formula I and formula II) 2.2.4 Exclusion criteria: Patients with systemic disease and patients with blood disease 2.3 STUDY METHOD 33 patients tested for the second First 2.3.1.test: Study design time (after 1-month treated anti-Prospective Peripheral blood count studies, interventions, described with vertical monitoring TB drugs) - Bone marrow aspirate 2.3.2 Sample selection for study - Peripheral blood count - Coagulation - Bonerepresentative marrow aspiratecriteria of Sample size: Applying the standard and important - Iron metabolism - Coagulation patients with pulmonary tuberculosis is anemia; calculated according to the - Iron metabolism - Immunoglobulin formula: - Immunoglobulin Analysis related to - Evaluate p.qall indices hematological after one 2month of n= α indices with: 111 1− treatment d patients new PTB - Compare with the and 47 patients with first indices We calculate n≥135PTB treated z Object 2: Object 1: 2.3.3 of STUDY DESIGN DIAGRAM Study some characteristics Evaluation of changes in peripheral blood and bone hematological indices in patients marrow testing in patients with with PTB before and after antinon-drug-resistant pulmonary tuberculosis drugs treatment tuberculosis treated at National Lung Hospital from 2015 to 2017 Chapter 3: RESULTS 3.1 Characteristics of hematological indices 3.1.1 Average value of some hematological indices a Average values of red blood cell The results on changes in peripheral red blood cells indices in patients with pulmonary tuberculosis are presented in Table 3.1, Chart 3.1: Table 3.1 Characteristics of erythrocyte indices of studied patients (n = 158) Patients Indices Min Max ( ± SD) Male 3.99 ± 0,92 1.87 6.32 Red blood cell (x 1012/l) Female 3.91 ± 0.75 2.33 5.45 112.4 ± 25.4 58 172 Hemoglobin (Hb) Male (g/l) Female 106 ± 23.3 68 156 Male 34.09 ± 7.11 18.8 49.5 Hct (Hematocrit) (%) Female 33.14 ± 6.45 20.1 45.9 MCV (fl) 86.32 ± 10.46 55.2 121 MCH (pg) 28.24 ± 3.61 18.4 36.4 MCHC (g/l) 327.1 ± 17.3 255 391 Reticulocyte 0.073 ± 0.081 0.01 0.86 Table 3.1 shows that the patients had a red blood cell count, both hemoglobin and hematocrit levels decreased Chart 3.1: Anemia characteristics of patients with PTB by sex The rate of mild anemia is higher in men than women and the rate of severe anemia is higher in women than men, the difference is statistically significant with p 500ng/ml) 109 68.99 3406.8±4979.94 Incrase Fibrinogen (>4g/l) 94 59.49 5.66±1.28 Incrase rTT (>1.25) 40 25.32 1.45±0.29 Incrase rAPTT (>1.25) 28 17.72 1.43±0.14 Decrase PT (